Abstract: The tRNA-derived fragments (tRF and tiRNA) are a newly discovered type of non-coding RNA (ncRNA) that has been found to be stably expressed in peripheral blood. Studies have shown that tRF and tiRNA play important roles in human tumors by regulating multiple processes, including gene expression and silencing, cell proliferation and apoptosis, and protein translation. The tissue-specificity, high abundance, and stability of tRF and tiRNA, along with their broad-spectrum functional roles, confer them significant advantages for use in the field of oncology research. There is increasing evidence that aberrantly expressed tRF and tiRNA may be potential biomarkers or therapeutic targets for tumor diagnosis and prognosis. This paper summarizes the source, structure, biological characteristics, and functions of different tRF and tiRNA subtypes and explores their potential relationship with tumors and their underlying mechanisms in order to provide a novel idea for the early diagnosis and targeted therapy of tumors.